SOLUBILITY PROPERTIES OF NEURONAL TUBULIN - EVIDENCE FOR LABILE AND STABLE MICROTUBULES
SOLUBILITY PROPERTIES OF NEURONAL TUBULIN - EVIDENCE FOR LABILE AND STABLE MICROTUBULES
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DOI：
10.1016/0006-8993(84)90974-0
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发表时间：
1984-01-01
期刊：
影响因子：
2.9
通讯作者：
KURDYLA, JT
中科院分区：
文献类型：
--
作者：
BLACK, MM;COCHRAN, JM;KURDYLA, JT
用电镜和生化技术研究了大鼠交感神经元微管蛋白和微管的溶解特性。对于形态学分析，在存在或不存在1 mM CaCl 2的情况下用Triton X-100提取神经元，并检查所得洗涤剂提取的残留物的微管。在平行实验中，在各种溶液条件下测定微管蛋白的溶解度。洗涤剂提取的残留物的神经元制备无钙含有许多微管。在存在Ca 2+的情况下制备的神经突残余物也含有微管，但数量远低于在不存在Ca 2+的情况下制备的残余物。生化数据与形态学观察结果一致。在微管稳定条件下进行去污剂提取后，30%的微管蛋白是去污剂可溶的（即，未聚合的），而70%是洗涤剂不溶性的（即，聚合的）。对微管蛋白的溶解度性质的更详细的检查表明，62%是洗涤剂不溶性的，但可溶于含有mM CaCl 2的缓冲液中，而5-8%是洗涤剂和Ca 2+不溶性的。非特异性吸附的微管蛋白到洗涤剂不溶性成分的文化，组装微管蛋白到预先存在的微管，和不完全提取的微管蛋白从细胞中的贡献最小的钙离子可溶性和不溶性微管蛋白的水平与所使用的提取条件。大多数神经元微管蛋白组装成微管，其在用Ca 2+处理后解聚。神经元微管蛋白的一部分组装成微管，微管显示出独特的Ca ~（2+）稳定性。
The solubility properties of tubulin and microtubules in pure cultures of rat sympathetic neurons were examined by EM and biochemical techniques. For morphological analyses, neurons were extracted with Triton X-100 in the presence or absence of 1 mM CaCl2, and the resulting detergent-extracted residues were examined for microtubules. In parallel experiments, the solubility of tubulin was determined under various solution conditions. Detergent-extracted residues of neurons prepared without Ca2+ contained many microtubules. Neurite residues prepared in the presence of Ca2+ also contained microtubules, but at substantially lower numbers than in residues prepared without Ca2+. The biochemical data parallel the morphological observations. Following detergent-extraction under microtubule stabilizing conditions, 30% of the tubulin was detergent-soluble (i.e., unpolymerized), while 70% was detergent-insoluble (i.e., polymerized). A more detailed examination of the solubility properties of tubulin indicated that 62% was detergent-insoluble but soluble in buffers containing mM CaCl2, while 5-8% was detergent and Ca2+-insoluble. Non-specific adsorption of tubulin onto detergent-insoluble components of the cultures, assembly of tubulin onto pre-existing microtubules, and incomplete extraction of tubulin from cells contributed minimally to the levels of Ca2+- soluble and insoluble tubulin obtained with the extraction conditions used. The majority of neuronal tubulin is assembled into microtubules which diassemble upon treatment with Ca2+. A portion of the neuronal tubulin is assembled into microtubules which show the unusal property of Ca2+-stability.